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破伤风类毒素免疫定量检测

Quantitative immunoassay detection of tetanus toxoid

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【作者】 钱雯黄微代云波施競李作生马波

【Author】 QIAN Wen;HUANG Wei;DAI Yun-bo;SHI Jing;LI Zuo-sheng;MA Bo;Yunnan Walvax Biotechnology Co.,LTD.;

【机构】 云南沃森生物技术股份有限公司

【摘要】 目的制备破伤风类毒素的单克隆抗体并建立竞争ELISA和双抗体夹心ELISA定量检测方法。方法用破伤风类毒素免疫小鼠,通过常规方法融合、间接ELISA筛选,有限稀释法克隆,制备破伤风类毒素的单克隆抗体。以制备的单克隆抗体进行竞争ELISA和双抗体夹心ELISA定量检测破伤风类毒素。结果应用制备的单抗进行的定量检测范围为0.003 2μg/ml~50μg/ml。2种方法对同一未知浓度样品的破伤风类毒素的检测结果分别为0.045 89μg/ml和0.045 94μg/ml。结论用制备的抗破伤风类毒素的单克隆抗体,建立了破伤风类毒素ELISA定量检测方法。

【Abstract】 Objective To prepare monoclonal antibodies against tetanus toxoid and establish competitive ELISA and double antibody sandwich ELISA methods for the quantitative detection of tetanus toxoid. Methods Mice immunized with tetanus toxoid,and with conventional methods for fusion,indirect ELISA for screening,limiting dilution method for cloning,to prepare monoclonal antibodies against tetanus toxoid. The prepared monoclonal antibody was used to detect the tetanus toxoid with quantitatively competing against ELISA and double antibody sandwich ELISA. Results The detection range of quantitative detection of tetanus toxin was 0. 003 2 μg / ml-50 μg / ml with prepared monoclonal antibody. The results detected from the two methods for contents of same sample of tetanus toxin unknown concentration were 0. 045 89 μg / ml and 0. 045 94 μg / ml,respectively. Conclusion The monoclonal antibodies against tetanus toxoid were prepared and ELISA quantitative detection methods for tetanus toxoid were established.

【基金】 国家科技重大专项重大新药创制专项“创新性吸附无细胞百白破3组分联合疫苗的研究开发”(2013ZX-09102036)
  • 【文献出处】 中国卫生检验杂志 ,Chinese Journal of Health Laboratory Technology , 编辑部邮箱 ,2015年07期
  • 【分类号】R392
  • 【被引频次】1
  • 【下载频次】215
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